MiRNA expression in primary human leukaemic B-cells from peripheral blood and after culture on stromal cells in the presence and absence of CD154 and IL-4
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE24694 | GEO | 2011/11/03
SECONDARY ACCESSION(S): PRJNA132267
REPOSITORIES: GEO
Similar Datasets
Project description:Dynamic miRNA expression data in 377 miRNA present on Taqman low density array (TLDA) Human MicroRNA Array (A) plates (Applied Biosystems, #4398965). Primary human leukaemic B-cells were cultured for 24 hours on a stromal cell layer or a stromal cell layer with CD154 and IL-4 in order to find out how miRNA expression compared to that of freshly isolated leukaemic cells.
2011-11-03 | E-GEOD-24694 | biostudies-arrayexpress
Project description:Exosomes are secreted into the intercellular space and carry a diverse cargo of proteins, mRNA and miRNA that have important roles in cell-to-cell communication. We found that exosomes from primary human leukaemic B-cells contain a population of miRNAs that has been selected from the general cellular pool to be enriched in miR-323-3p, miR-374b, miR-363 and miR-494. Gene targets of these miRNA are found within the TGFβ pathway, but we noted that CD69, a protein that is essential for the regulation of normal lymphocyte egress from lymph nodes was the top predicted target of miR-363. Utilising luciferase reporter assays we demonstrate that CD69 is a target of miR-363. The interaction between miR-363 and CD69 is functionally important as demonstrated by repression of CD69 expression by a miR-363 mimic in the Jurkat T-cell line. MicroRNA expression profiles of isolated exosomes and CLL cells cultured on CD154/IL-4 feeder layers for 24 hours were determined using Taqman low-density miRNA (TLDA) arrays. Patient samples were layered on Ficoll and centrifuged for 20 minutes at 2500 rpm following which the interface layer conatining mononuclear cells was removed. The cells were washed in PBS and cultured on a feeder layer of mouse fibroblasts transfected with human CD154 in the presence of IL-4 (10 ng/ml) for 24 hours MiRNA extracts were made using a miRvana miRNA isolation kit (Applied Biosystems, Paisley, UK), and reverse transcribed using MegaPlex RT Primers and a TaqMan MicroRNA Reverse Transcription Kit (Applied Biosystems). Equal amounts of RNA were added to each microarray plate. Human microRNA Array (A) plates (containing assays for 377 miRNA) were used (Applied Biosystems, #4398965) and reactions carried out on a 7900HT Real Time PCR system.
2014-07-25 | GSE54434 | GEO
Project description:Exosomes are secreted into the intercellular space and carry a diverse cargo of proteins, mRNA and miRNA that have important roles in cell-to-cell communication. We found that exosomes from primary human leukaemic B-cells contain a population of miRNAs that has been selected from the general cellular pool to be enriched in miR-323-3p, miR-374b, miR-363 and miR-494. Gene targets of these miRNA are found within the TGFM-NM-2 pathway, but we noted that CD69, a protein that is essential for the regulation of normal lymphocyte egress from lymph nodes was the top predicted target of miR-363. Utilising luciferase reporter assays we demonstrate that CD69 is a target of miR-363. The interaction between miR-363 and CD69 is functionally important as demonstrated by repression of CD69 expression by a miR-363 mimic in the Jurkat T-cell line. MicroRNA expression profiles of isolated exosomes and CLL cells cultured on CD154/IL-4 feeder layers for 24 hours were determined using Taqman low-density miRNA (TLDA) arrays. Patient samples were layered on Ficoll and centrifuged for 20 minutes at 2500 rpm following which the interface layer conatining mononuclear cells was removed. The cells were washed in PBS and cultured on a feeder layer of mouse fibroblasts transfected with human CD154 in the presence of IL-4 (10 ng/ml) for 24 hours MiRNA extracts were made using a miRvana miRNA isolation kit (Applied Biosystems, Paisley, UK), and reverse transcribed using MegaPlex RT Primers and a TaqMan MicroRNA Reverse Transcription Kit (Applied Biosystems). Equal amounts of RNA were added to each microarray plate. Human microRNA Array (A) plates (containing assays for 377 miRNA) were used (Applied Biosystems, #4398965) and reactions carried out on a 7900HT Real Time PCR system. MiRNA expression was compared in paired samples of chronic lymphocytic leukaemia cells and exosomes utilising Taqman Real-Time PCR.
2014-07-25 | E-GEOD-54434 | biostudies-arrayexpress
Project description:Aim of this study was to identify miRNAs which are regulated by LIF in different trophoblastic cell lines Total RNA was reverse-transcribed, pre-amplified with miRNA-specific primers, and loaded into TaqMan Arrays. Quantitative real-time PCR was performed for 768 miRNAs. For each of two cards per sample (A and B), 9 ul of pre-amplified diluted sample was mixed with TaqMan reaction mix (Applied Biosystems) and loaded into miRNA TLDA cards (Applied Biosystems) by centrifugation. Cards were sealed, and qRT-PCR was performed with a real-time thermocycler (ABI-7900, Applied Biosystems) per manufacturer's recommendations.
2019-09-01 | GSE130489 | GEO
Project description:To gain more insight in the biological expression and role of microRNAs in trophoblast cells, we then set up to determinate the miRNA expression pattern of four trophoblastic cell lines and compare them with the miRNA signature of isolated trophoblast Total RNA was reverse-transcribed, pre-amplified with miRNA-specific primers, and loaded into TLDA cards. Quantitative real-time PCR was performed for 768 miRNAs. For each of two cards per sample (A and B), 9 ul of pre-amplified diluted sample was mixed with TaqMan reaction mix (Applied Biosystems) and loaded into miRNA TLDA cards (Applied Biosystems) by centrifugation. Cards were sealed, and qRT-PCR was performed with a real-time thermocycler (ABI-7900, Applied Biosystems) per manufacturer's recommendations.
2011-09-23 | GSE32346 | GEO
Project description:We determined expression profiles of 667 miRNAs using TaqMan Low Density Arrays (TLDA-TaqMan Array Human MicroRNA Card Set v2.0, Applied Biosystems) in 8 samples of colorectal cancer tissues and 8 samples of paired non-tumoral colonic tissues.
2012-03-31 | E-MEXP-3515 | biostudies-arrayexpress
Project description:Eosinophlic esophagitis (EoE) is increasely recognized as an antigen-drived disorder. The goal of this study is to reveal the miRNA expression changes in EoE before and after a successful glucocorticoid steroid treatment. Total RNA was extracted from the esophageal epithelial layers of 5 paired paraffin-embedded biopsies before and after treatment with glucocorticosteroids using RecoverAll Total Nucleic Acid Extraction Kit for FFPE tissues (Ambion, Austin, TX). Five nanograms of total RNA was reverse-transcribed using the Taqman MicroRNA Reverse Transcription Kit and the Megaplex RT primer Human Pool A (Applied Biosystems). The reverse-transcribed cDNA was then pre-amplified in 12 cycles of PCR using Taqman PreAmp Master Mix and the Megaplex PreAmp primers, Human Pool A (Applied Biosystems). The cDNA’s were then diluted and loaded on to a Taqman Human miRNA Array card A (Platform GPL9731 ; Applied Biosystems), which contains probes for 377 distinct miRNAs. The Array cards were run on an ABI HT7900 qPCR instrument. Ct values were obtained for all miRNAs represented on the cards and fold changes in expression were calculated using the delta delta Ct (ddCt) method.
2012-03-23 | E-GEOD-36726 | biostudies-arrayexpress
Project description:VSMCs were cultured on Jag1 or IgG-coated plates as described above. RNA was extracted using RNA isolation kit (Qiagen). Reverse-transcription PCR was done with RNA-to-cDNA kit (Applied Biosystems). cDNA was run on a TaqMan Low-Density Array, human angiogenesis panel (Applied Biosystems , Cat#4378725), amplified on a 7900 HT Fast Real Time PCR system (Applied Biosystems) according to manufacturer’s instructions.
2011-12-01 | GSE34024 | GEO
Project description:To gain more insight in the expression and role of microRNAs in trophoblastic cells, we have assessed the miRNA expression pattern of four trophoblastic cell lines and have compared them with the miRNA signature of human isolated 3rd trimester cytotrophoblast cells. Total RNA was reverse-transcribed, pre-amplified with miRNA-specific primers, and loaded into TLDA cards. Quantitative real-time PCR was performed for 768 miRNAs. For each of two cards per sample (A and B), 9 ul of pre-amplified diluted sample was mixed with TaqMan reaction mix (Applied Biosystems) and loaded into miRNA TLDA cards (Applied Biosystems) by centrifugation. Cards were sealed, and qRT-PCR was performed at a real-time thermocycler (ABI-7900, Applied Biosystems) following the manufacturer's recommendations. MiRNA signatures of human primary 3rd trimester trophoblast cells were compared with those of four trophoblastic cell lines
2011-09-22 | E-GEOD-32346 | biostudies-arrayexpress
Project description:We aimed to identify urinary exosomal miRNAs associated with PCa metastasis and develop a non-invasive risk-scoring model for PCa metastasis in this study. MiRNA profiles were examined using the Taqman low-density miRNA array (TLDA). Megaplex reverse transcription reactions and pre-amplification reactions were performed to increase the quantity of cDNA for miRNA expression analysis using the Megaplex PreAmp Primers Human Pool A and TaqMan PreAmp Master Mix (Thermo Fisher Scientific). MiRNA expression was evaluated via the TLDA panel A v2.0 (Thermo Fisher Scientific). Raw data were processed using the QuantStudio Real-Time PCR Software (Thermo Fisher Scientific) to determine a cycle threshold (Ct) value for each miRNA.
2021-06-07 | GSE173094 | GEO